Rose Bengal Photothrombosis by Confocal Optical Imaging <em>In Vivo</em>: A Model of Single Vessel Stroke

Abstract

In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to image cellular events in an intact animal. Additionally, the ability to induce physiological disease states such as stroke in vivo increases its utility. The technique described herein allows for physiological assessment of cellular responses within the CNS following a stroke and can be adapted for other pathological conditions being studied. The technique presented uses laser excitation of the photosensitive dye Rose Bengal in vivo to induce a focal ischemic event in a single blood vessel.The video protocol demonstrates the preparation of a thin-skulled cranial window over the somatosensory cortex in a mouse for the induction of a Rose Bengal photothrombotic event keeping injury to the underlying dura matter and brain at a minimum. Surgical preparation is initially performed under a dissecting microscope with a custom-made surgical/imaging platform, which is then transferred to a confocal microscope equipped with an inverted objective adaptor. Representative images acquired utilizing this protocol are presented as well as time-lapse sequences of stroke induction. This technique is powerful in that the same area can be imaged repeatedly on subsequent days facilitating longitudinal in vivo studies of pathological processes following stroke. Video LinkThe video component of this article can be found at

Astrocyte plasticity in mice ensures continued endfoot coverage of cerebral blood vessels following injury and declines with age

“…Successful clot formation can be visualized by a dark area forming in the vessel, indicative of red blood cell accumulation, and intense fluorescence due to dye accumulation above that dark mass (Fig. 7d–f ) 53 . Critically, this method allowed us to focally and transiently occlude penetrating arterioles and thereby examine if a structural plasticity response would occur following focal loss of astrocyte-vascular coverage.…”

Quantitative behavioral evaluation of a non-human primate stroke model using a new monitoring system

“…This was confirmed by MRI-based imaging evaluation and phenotypic assessment using MarmoDetector. In recent years, the review ( Uzdensky, 2018 ) and excellent practical guidance ( Labat-gest and Tomasi, 2013 ; Talley Watts et al, 2015 ; Song et al, 2016 ) have been published on photothrombotic stroke models. Photothrombotic stroke causes endothelial damage and platelet aggregation in vessels, resulting in distinct ischemic lesions in the cerebral cortex ( Watson et al, 1985 ).…”

Understanding the Pathophysiology of Thrombotic APS through Animal Models

“…Rose Bengal is a photosensitizing fluorescent dye that produces reactive oxygen species and focal vascular endothelial damage following exposure to green light. Beyond hamsters, the photochemical carotid model has been commonly used in vascular studies of thrombosis in other rodent species, including mice, although not to our knowledge in the context of APS [ 53 , 54 , 55 ].…”